Cytotoxicity Screening of Endophytic Fungi from Phaleria macrocarpa (Scheff) Boerl) Collected in Yogyakarta District
Keywords:
Phaleria macrocarpa, cytotoxicity, endophyte, cancer
Abstract
Cancer is a disease that is a global problem with a high prevalence and mortality rate. Indonesia has biodiversity with enormous potential as a source of medicinal ingredients. Phaleria macrocarpa (Scheff) Boerl) It is known to contain active metabolites, potentially an anticancer. Some endophytes are reported to be able to produce secondary metabolites similar to their parent plants. This study aimed to obtain endophytic fungi from P. macrocarpa capable of producing compounds with anticancer potential. Endophytes producing anticancer compounds were explored by screening endophytic fungi producing secondary metabolites from parts of the plant P. macrocarpa collected in the Yogyakarta area. The anticancer bioassay (MTT assay) was performed in vitro using cancer cells T47D, MCF7, HeLa, and normal Vero cells. Potential extracts were traced for their mechanism of cell cycle modulation and apoptosis using Flowcytometry. First, fungal identification was carried out on fungi producing potential cytotoxic extracts. Then the compound content was analyzed using LC-HRMS. The results from the six extracts obtained from fruit, leaf, and stem parts showed that the endophytic extract from the leaves (code PC-L2) had the best cytotoxicity compared to other endophytes and showed the best result on MCF-7 cells with an IC50 value of 110.66 µg/mL. The producing PC-L2 extract fungi are closely associated with Clonostachys wenpingii strain DUCC5606. This extract induced apoptosis but not through the mechanism of cell cycle modulation.
References
Abdel-Wahab, N. M., Harwoko, H., Mueller, W. E., Hamacher, A., Kassack, M. U., Fouad, M. A., Kamel, M.S.; Lin, W.; Ebrahim, W.; Liu, Z & Proksch, P. (2019). Cyclic heptapeptides from the soil-derived fungus Clonostachys rosea. Bioorganic & Medicinal Chemistry, 27(17), 3954-3959. https://doi.org/10.1016/j.bmc.2019.07.025
Amir, H., Murcitro, B. G., Ahmad, A. S., & Kassim, M. N. I. (2017). The potential use of Phaleria macrocarpa leaves extract as an alternative drug for breast cancer among women living in poverty. Asian Journal For Poverty Studies (AJPS), 3(2), 138-145.
Ashoka, H., Hegde, P., Manasa, K. H., Madihalli, C., Pradeep, S., & Shettihalli, A. K. (2017). Isolation and detection of vinca alkaloids from endophytes isolated from Catharanthus roseus. Eur. J. Biomed. Pharm. Sci, 10, 675-683.
Astuti, P., Januarti, I. B., Kiromah, N. Z. W., Fitri, H. A., Wahyono, W., & Wahyuono, S. (2020). Pyrophen Isolated from the Endophytic Fungus Aspergillus fumigatus Strain KARSV04 Synergizes the Effect of Doxorubicin in Killing MCF7 but not T47D Cells. Turkish Journal of Pharmaceutical Sciences, 17(3), 280. https://10.4274/tjps.galenos.2019.30633
Astuti, P., Nuryastuti, T., & Purwantini, I. (2014). Antimicrobial and Cytotoxic Activities of Endophytic Fungi Isolated from Artemisia annua L. Journal of Applied Pharmaceutical Science, 4(10): 047-050.
Barreto, R., Waning, D. L., Gao, H., Liu, Y., Zimmers, T. A., & Bonetto, A. (2016). Chemotherapy-related cachexia is associated with mitochondrial depletion and the activation of ERK1/2 and p38 MAPKs. Oncotarget, 7(28).
Bézivin, C., Tomasi, S., Lohézic-Le Dévéhat, F., & Boustie, J. (2003). Cytotoxic activity of some lichen extracts on murine and human cancer cell lines. Phytomedicine, 10(6-7), 499-503.
Cheplick, G. P., Faeth, S., & Faeth, S. H. (2009). Ecology and evolution of the grass-endophyte symbiosis. OUP USA.
Chutulo, E. C., & Chalannavar, R. K. (2018). Endophytic mycoflora and their bioactive compounds from Azadirachta indica: A comprehensive review. Journal of Fungi, 4(2), 42. https://doi.org/10.3390/jof4020042
Ferlay, J., Ervik, M., Lam, F., Colombet, M., Mery, L., Piñeros, M., A. Znaor, I., Soerjomataram & Bray, F. (2020). Global Cancer Observatory: Cancer Today. Lyon: International Agency for Research on Cancer. Retrieved from https://gco.iarc.fr/today, avaliable February 2021).
Gasong, B. T., & Tjandrawinata, R. R. (2016). Production of secondary metabolite E2. 2 from Phaleria macrocarpa endophytic fungus. Asian Pacific Journal of Tropical Biomedicine, 6(10), 881-885. https://doi.org/10.1016/j.apjtb.2016.01.005
Hain, B. A., Jude, B., Xu, H., Smuin, D. M., Fox, E. J., Elfar, J. C., & Waning, D. L. (2020). Zoledronic acid improves muscle function in healthy mice treated with chemotherapy. Journal of Bone and Mineral Research, 35(2), 368-381. https://doi.org/10.1002/jbmr.3890
Han, P., Zhang, X., Xu, D., Zhang, B., Lai, D., & Zhou, L. (2020). Metabolites from Clonostachys fungi and their biological activities. Journal of fungi, 6(4), 229. https://doi.org/10.3390/jof6040229
Kusari, S., Pandey, S. P., & Spiteller, M. (2013). Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites. Phytochemistry, 91, 81-87. https://doi.org/10.1016/j.phytochem.2012.07.021
Lesmana, D., Andrianto, D., & Astuti, R. I. (2021). Antiaging Properties of the Ethanol Fractions of Clove (Syzygium aromaticum L.) Bud and Leaf at the Cellular Levels: Study in Yeast Schizosaccharomyces pombe. Scientia Pharmaceutica, 89(4), 45. https://doi.org/10.3390/scipharm89040045
Li, E., Hou, B., Gao, Q., Xu, Y., Zhang, C., Liu, X., Jiang, X., & Che, Y. (2020). Disulfide cleavage in a dimeric epipolythiodioxopiperazine natural product diminishes its apoptosis-inducing effect but enhances autophagy in tumor cells. Journal of natural products, 83(3), 601-609. https://doi.org/10.1021/acs.jnatprod.9b00994
Liu, L., Liu, Y., Zhao, J., Xing, X., Zhang, C., & Meng, H. (2020). Neuroprotective effects of D-(-)-quinic acid on aluminum chloride-induced dementia in rats. Evidence-Based Complementary and Alternative Medicine. https://doi.org/10.1155/2020/5602597
Padamsee, T. J., Wills, C. E., Yee, L. D., & Paskett, E. D. (2017). Decision making for breast cancer prevention among women at elevated risk. Breast Cancer Research, 19(1), 1-12. https://doi.org/10.1186/s13058-017-0826-5
Pero, R. W., Lund, H., & Leanderson, T. (2009). Antioxidant metabolism induced by quinic acid. Increased urinary excretion of tryptophan and nicotinamide. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 23(3), 335-346.
Pirttilä, A. M., & Frank, A. C. (2011). Endophytes of forest trees. Springer Netherlands.
Shwter, A. N., Abdullah, N. A., Alshawsh, M. A., El-Seedi, H. R., Al-Henhena, N. A., Khalifa, S. A., & Abdulla, M. A. (2016). Chemopreventive effect of Phaleria macrocarpa on colorectal cancer aberrant crypt foci in vivo. Journal of ethnopharmacology, 193, 195-206. https://doi.org/10.1016/j.jep.2016.08.002
Singh, N., Baby, D., Rajguru, J. P., Patil, P. B., Thakkannavar, S. S., & Pujari, V. B. (2019). Inflammation and cancer. Annals of African medicine, 18(3), 121. https://doi.org/10.4103/aam.aam_56_18
Syarmalina, S., Lely, W., & Laupa, N. (2007). Uji Sitotoksik Hasil Fermentasi Kapang Endofit Buah Mahkota Dewa (Phaleria macrocarpa) terhadap Sel MCF-7. Jurnal Ilmu Kefarmasian Indonesia, 5(1), 23-30.
Tan, R. X., & Zou, W. X. (2001). Endophytes: a rich source of functional metabolites. Natural product reports, 18(4), 448-459. DOI https://doi.org/10.1039/B100918O
Tanaka, S., Kawashima, K., Usami, M., Nakaura, S., Kodama, Y., & Takanaka, A. (1990). Studies on the teratogenic potential of 2, 2'-methylenebis (4-methyl-6-tert-butylphenol) in rats. Eisei Shikenjo hokoku. Bulletin of National Institute of Hygienic Sciences, (108), 52-57.
Tandrasasmita, O. M., Lee, J. S., Baek, S. H., & Tjandrawinata, R. R. (2010). Induction of cellular apoptosis in human breast cancer by DLBS1425, a Phaleria macrocarpa compound extract, via down-regulation of PI3-kinase/AKT pathway. Cancer biology & therapy, 10(8), 814-823. https://doi.org/10.4161/cbt.10.8.13085
Todoric, J., Antonucci, L., & Karin, M. (2016). Targeting inflammation in cancer prevention and therapy. Cancer Prevention Research, 9(12), 895-905. 10.1158/1940-6207.CAPR-16-0209
Vijaya, T. (2017). Anticancer, antimicrobial and antioxidant bioactive factors derived from marine fungal endophytes; a review. Indo Am. J. Pharm. Res, 7, 7313-7321.
Weerapreeyakul, N., Nonpunya, A., Barusrux, S., Thitimetharoch, T., & Sripanidkulchai, B. (2012). Evaluation of the anticancer potential of six herbs against a hepatoma cell line. Chinese medicine, 7(1), 1-7.
Zeng, Z. Q., & Zhuang, W. Y. (2022). Three New Species of Clonostachys (Hypocreales, Ascomycota) from China. Journal of Fungi, 8(10), 1027. https://doi.org/10.3390/jof8101027
Zhang, S. Y., Zhang, Q. H., Zhao, W., Zhang, X., Zhang, Q., Bi, Y. F., & Zhang, Y. B. (2012). Isolation, characterization and cytotoxic activity of benzophenone glucopyranosides from Mahkota Dewa (Phaleria macrocarpa (Scheff.) Boerl). Bioorganic & medicinal chemistry letters, 22(22), 6862-6866. https://doi.org/10.1016/j.bmcl.2012.09.038
Zheng, Z., Wang, Y., Yu, H., Li, W., Wu, J., Cai, C., & He, Y. (2020). Salvianolic acid B inhibits ototoxic drug–induced ototoxicity by suppression of the mitochondrial apoptosis pathway. Journal of Cellular and Molecular Medicine, 24(12), 6883-6897. https://doi.org/10.1111/jcmm.15345
Amir, H., Murcitro, B. G., Ahmad, A. S., & Kassim, M. N. I. (2017). The potential use of Phaleria macrocarpa leaves extract as an alternative drug for breast cancer among women living in poverty. Asian Journal For Poverty Studies (AJPS), 3(2), 138-145.
Ashoka, H., Hegde, P., Manasa, K. H., Madihalli, C., Pradeep, S., & Shettihalli, A. K. (2017). Isolation and detection of vinca alkaloids from endophytes isolated from Catharanthus roseus. Eur. J. Biomed. Pharm. Sci, 10, 675-683.
Astuti, P., Januarti, I. B., Kiromah, N. Z. W., Fitri, H. A., Wahyono, W., & Wahyuono, S. (2020). Pyrophen Isolated from the Endophytic Fungus Aspergillus fumigatus Strain KARSV04 Synergizes the Effect of Doxorubicin in Killing MCF7 but not T47D Cells. Turkish Journal of Pharmaceutical Sciences, 17(3), 280. https://10.4274/tjps.galenos.2019.30633
Astuti, P., Nuryastuti, T., & Purwantini, I. (2014). Antimicrobial and Cytotoxic Activities of Endophytic Fungi Isolated from Artemisia annua L. Journal of Applied Pharmaceutical Science, 4(10): 047-050.
Barreto, R., Waning, D. L., Gao, H., Liu, Y., Zimmers, T. A., & Bonetto, A. (2016). Chemotherapy-related cachexia is associated with mitochondrial depletion and the activation of ERK1/2 and p38 MAPKs. Oncotarget, 7(28).
Bézivin, C., Tomasi, S., Lohézic-Le Dévéhat, F., & Boustie, J. (2003). Cytotoxic activity of some lichen extracts on murine and human cancer cell lines. Phytomedicine, 10(6-7), 499-503.
Cheplick, G. P., Faeth, S., & Faeth, S. H. (2009). Ecology and evolution of the grass-endophyte symbiosis. OUP USA.
Chutulo, E. C., & Chalannavar, R. K. (2018). Endophytic mycoflora and their bioactive compounds from Azadirachta indica: A comprehensive review. Journal of Fungi, 4(2), 42. https://doi.org/10.3390/jof4020042
Ferlay, J., Ervik, M., Lam, F., Colombet, M., Mery, L., Piñeros, M., A. Znaor, I., Soerjomataram & Bray, F. (2020). Global Cancer Observatory: Cancer Today. Lyon: International Agency for Research on Cancer. Retrieved from https://gco.iarc.fr/today, avaliable February 2021).
Gasong, B. T., & Tjandrawinata, R. R. (2016). Production of secondary metabolite E2. 2 from Phaleria macrocarpa endophytic fungus. Asian Pacific Journal of Tropical Biomedicine, 6(10), 881-885. https://doi.org/10.1016/j.apjtb.2016.01.005
Hain, B. A., Jude, B., Xu, H., Smuin, D. M., Fox, E. J., Elfar, J. C., & Waning, D. L. (2020). Zoledronic acid improves muscle function in healthy mice treated with chemotherapy. Journal of Bone and Mineral Research, 35(2), 368-381. https://doi.org/10.1002/jbmr.3890
Han, P., Zhang, X., Xu, D., Zhang, B., Lai, D., & Zhou, L. (2020). Metabolites from Clonostachys fungi and their biological activities. Journal of fungi, 6(4), 229. https://doi.org/10.3390/jof6040229
Kusari, S., Pandey, S. P., & Spiteller, M. (2013). Untapped mutualistic paradigms linking host plant and endophytic fungal production of similar bioactive secondary metabolites. Phytochemistry, 91, 81-87. https://doi.org/10.1016/j.phytochem.2012.07.021
Lesmana, D., Andrianto, D., & Astuti, R. I. (2021). Antiaging Properties of the Ethanol Fractions of Clove (Syzygium aromaticum L.) Bud and Leaf at the Cellular Levels: Study in Yeast Schizosaccharomyces pombe. Scientia Pharmaceutica, 89(4), 45. https://doi.org/10.3390/scipharm89040045
Li, E., Hou, B., Gao, Q., Xu, Y., Zhang, C., Liu, X., Jiang, X., & Che, Y. (2020). Disulfide cleavage in a dimeric epipolythiodioxopiperazine natural product diminishes its apoptosis-inducing effect but enhances autophagy in tumor cells. Journal of natural products, 83(3), 601-609. https://doi.org/10.1021/acs.jnatprod.9b00994
Liu, L., Liu, Y., Zhao, J., Xing, X., Zhang, C., & Meng, H. (2020). Neuroprotective effects of D-(-)-quinic acid on aluminum chloride-induced dementia in rats. Evidence-Based Complementary and Alternative Medicine. https://doi.org/10.1155/2020/5602597
Padamsee, T. J., Wills, C. E., Yee, L. D., & Paskett, E. D. (2017). Decision making for breast cancer prevention among women at elevated risk. Breast Cancer Research, 19(1), 1-12. https://doi.org/10.1186/s13058-017-0826-5
Pero, R. W., Lund, H., & Leanderson, T. (2009). Antioxidant metabolism induced by quinic acid. Increased urinary excretion of tryptophan and nicotinamide. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 23(3), 335-346.
Pirttilä, A. M., & Frank, A. C. (2011). Endophytes of forest trees. Springer Netherlands.
Shwter, A. N., Abdullah, N. A., Alshawsh, M. A., El-Seedi, H. R., Al-Henhena, N. A., Khalifa, S. A., & Abdulla, M. A. (2016). Chemopreventive effect of Phaleria macrocarpa on colorectal cancer aberrant crypt foci in vivo. Journal of ethnopharmacology, 193, 195-206. https://doi.org/10.1016/j.jep.2016.08.002
Singh, N., Baby, D., Rajguru, J. P., Patil, P. B., Thakkannavar, S. S., & Pujari, V. B. (2019). Inflammation and cancer. Annals of African medicine, 18(3), 121. https://doi.org/10.4103/aam.aam_56_18
Syarmalina, S., Lely, W., & Laupa, N. (2007). Uji Sitotoksik Hasil Fermentasi Kapang Endofit Buah Mahkota Dewa (Phaleria macrocarpa) terhadap Sel MCF-7. Jurnal Ilmu Kefarmasian Indonesia, 5(1), 23-30.
Tan, R. X., & Zou, W. X. (2001). Endophytes: a rich source of functional metabolites. Natural product reports, 18(4), 448-459. DOI https://doi.org/10.1039/B100918O
Tanaka, S., Kawashima, K., Usami, M., Nakaura, S., Kodama, Y., & Takanaka, A. (1990). Studies on the teratogenic potential of 2, 2'-methylenebis (4-methyl-6-tert-butylphenol) in rats. Eisei Shikenjo hokoku. Bulletin of National Institute of Hygienic Sciences, (108), 52-57.
Tandrasasmita, O. M., Lee, J. S., Baek, S. H., & Tjandrawinata, R. R. (2010). Induction of cellular apoptosis in human breast cancer by DLBS1425, a Phaleria macrocarpa compound extract, via down-regulation of PI3-kinase/AKT pathway. Cancer biology & therapy, 10(8), 814-823. https://doi.org/10.4161/cbt.10.8.13085
Todoric, J., Antonucci, L., & Karin, M. (2016). Targeting inflammation in cancer prevention and therapy. Cancer Prevention Research, 9(12), 895-905. 10.1158/1940-6207.CAPR-16-0209
Vijaya, T. (2017). Anticancer, antimicrobial and antioxidant bioactive factors derived from marine fungal endophytes; a review. Indo Am. J. Pharm. Res, 7, 7313-7321.
Weerapreeyakul, N., Nonpunya, A., Barusrux, S., Thitimetharoch, T., & Sripanidkulchai, B. (2012). Evaluation of the anticancer potential of six herbs against a hepatoma cell line. Chinese medicine, 7(1), 1-7.
Zeng, Z. Q., & Zhuang, W. Y. (2022). Three New Species of Clonostachys (Hypocreales, Ascomycota) from China. Journal of Fungi, 8(10), 1027. https://doi.org/10.3390/jof8101027
Zhang, S. Y., Zhang, Q. H., Zhao, W., Zhang, X., Zhang, Q., Bi, Y. F., & Zhang, Y. B. (2012). Isolation, characterization and cytotoxic activity of benzophenone glucopyranosides from Mahkota Dewa (Phaleria macrocarpa (Scheff.) Boerl). Bioorganic & medicinal chemistry letters, 22(22), 6862-6866. https://doi.org/10.1016/j.bmcl.2012.09.038
Zheng, Z., Wang, Y., Yu, H., Li, W., Wu, J., Cai, C., & He, Y. (2020). Salvianolic acid B inhibits ototoxic drug–induced ototoxicity by suppression of the mitochondrial apoptosis pathway. Journal of Cellular and Molecular Medicine, 24(12), 6883-6897. https://doi.org/10.1111/jcmm.15345
Published
2023-09-26
How to Cite
Agustina, I., Lay, C. S., Astuti, P., & Hertiani, T. (2023). Cytotoxicity Screening of Endophytic Fungi from Phaleria macrocarpa (Scheff) Boerl) Collected in Yogyakarta District. Indonesian Journal of Pharmacy. https://doi.org/10.22146/ijp.7415
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Section
Research Article
